The continued and exciting development of next-generation sequencing (NGS) technologies harnesses considerable potential for the biotech industry. As NGS technologies advance, so do scientific breakthroughs. NGS allows scientists to better understand the human microbiome, uncover and analyze RNA variants, and identify rare variants in pathogens, cancers, and more. None of these innovations would be possible without the modern technology that powers next-generation sequencing. Current NGS technologies can sequence DNA faster and with higher sensitivity than ever before. As such, it’s apparent that having access to the appropriate NGS instrument is a crucial part of furthering research and applications of NGS. So how do you select or develop a technology that’s right for the job? Discover what to look for in NGS instruments with this guide.

An Introduction to Next-Generation Sequencing

Next-generation sequencing (NGS) is the latest generation of DNA and RNA sequencing, a process that allows scientists to determine the order of the bases within a DNA molecule. This in turn reveals genetic information about that specific DNA or RNA segment. Early methods of sequencing have been around since the 1970s, when techniques revolved around two-dimensional chromatography. In 1977, Frederick Sanger introduced the chain termination method, which offered the ability to sequence DNA reliably and in a reproducible method. These advancements paved the way for the first full DNA genome sequencing. In 1984, scientists used DNA sequencing to decipher the Epstein-Barr virus, marking a turning point—and indicating the potential of DNA sequencing.

In the late 1990s, new methods of DNA sequencing emerged. These new techniques and technologies had an incredibly high throughput compared to first-generation techniques. As such, they earned the name “next-generation sequencing.” NGS technology is highly scalable, which means it can sequence an entire genome at once by fragmenting it into smaller pieces that can be sampled simultaneously.

This parallel sequencing method has transformed DNA sequencing, bringing the world’s attention to NGS and the biotech industry. With such attention and potential, an increasing number of companies are looking for ways to leverage NGS. In order to do this, one must understand how the NGS workflow works and what role NGS technology plays in every step of the way.

The Next-Generation Sequencing Workflow

How does NGS actually work? The workflow involves four basic steps. Scientists begin with library preparation and cluster generation before moving onto the sequencing stage. Once sequencing is complete, experts can perform alignment and data analysis. Let’s delve deeper into each step of the NGS workflow and the technology involved throughout the process below:

Library preparation

NGS begins with library preparation. Specialists create a sequencing library by fragmenting samples into shorter sequences to prevent sequencing bias. Special adapter sequences are ligated to the ends of each fragment, which make the sequences compatible with the sequencer. Additional sequences, usually referred to as barcodes or indexes can also be added to the samples. These allow multiplexing of samples on the same sequencing run to save time and cost. . For RNA sequencing, RNA must first be converted to cDNA. The prepared library can undergo QC checks using real-time PCR to confirm DNA quality and quantity. This will allow the correct concentration of sample to be loaded onto the sequencer.

Cluster generation

The next step is to load the library into a flow cell. Here, each fragment undergoes amplification to create distinct, clonal clusters. This process is called cluster generation and creates millions of copies of single-stranded DNA. After cluster generation is complete, the samples are ready for sequencing.

Sequencing

DNA sequencing can be performed using several well-established methods: pyrosequencing, sequencing by synthesis (SBS), sequencing by ligation, or ion semiconductor sequencing. The platform selected will depend on the required chemistry, clonal amplification of library fragments either by emulsion PCR or on the sequencer itself (bridge PCR).

Data analysis

Analysis methods are dependent on the aim of the study. Many modern NGS instruments provide analysis tools that automate processes like sequence alignment or data visualization and interpretation. The generated data files are analyzed according to the workflow used.

A Guide to Next-Generation Sequencing Technology

The goal of NGS instruments is to increase throughput and offer scalable DNA sequencing solutions. These efficient and flexible systems make DNA sequencing more accessible for every kind of project and study. Scientists no longer need massive genome centers to do their work. As the technology becomes more accessible, new breakthroughs in research and novel applications will be uncovered. That’s why modern NGS technologies continue to push the boundaries of speed, flexibility, and scalability. These and other qualities make up what to look for in NGS instruments.

Considerations When Purchasing Next-Generation Sequencing Instruments

Like all devices, an NGS sequencer (and related equipment) must fit the needs of its end users. This means developers and buyers alike need to consider factors such as technical specifications, throughput requirements, price, size, and context of use. While sequencers are important, labs must also invest in high-quality equipment to support the overall NGS workflow. Nucleic acid quantity and quality analyzers, and thermocyclers are crucial in the library preparation step. Additionally, biotech cartridge engineering creates assay protocols and aids in executing techniques like PCR, DNA and RNA purification, and other crucial steps in NGS. If NGS technology is going to fulfill the needs of its users, it needs to incorporate all these devices in a manner that is efficient, reliable, and relatively easy to use.

Applications of Next-Generation Sequencing

NGS technology makes DNA sequencing faster, cheaper, and more reliable than ever. When researchers can easily compare large stretches of DNA from various individuals, they can dig deeper into genetic susceptibility or responses to diseases, environments, and other conditions. This information helps us better understand genetic conditions and elucidate new treatment regimes. NGS enables physicians to identify specific cancers in order to provide more appropriate treatments, perform preconception screening of couples planning on parenthood for heritable disorders, perform non-invasive prenatal testing (NIPT) to determine the risk that a fetus will be born with certain genetic abnormalities, and provide early detection in newborns to identify conditions that can affect a child’s long-term health and survival. All this progress has come in the last couple of decades with the rise of NGS technology. However, the methods and instruments behind NGS continue to evolve. As medical device developers expand on what their technology can do, researchers and scientists make new advancements in the biotech industry. The capacity of NGS to interrogate hundreds, thousands, and possibly millions of targets simultaneously offers incredible potential for a myriad of applications in healthcare settings. In order to serve the need for customized end-to-end solutions to meet the changing customer requirements, Gener8, is constantly looking for ways to improve our processes for mechanical engineering of medical devices and consumable products for biotech companies developing technologies such as NGS instruments. As the biotech market grows, your company faces fierce competition. Let Gener8 bring your ideas to market faster. With experts in science, engineering, and design, we have the necessary resources to create an expert device that fulfills your vision and meets the needs of its end users.